Topical nitric oxide systems and methods of use thereof

ABSTRACT

The present invention generally relates to compositions and methods for transdermal drug delivery. The compositions can be used in a variety of applications, including treating erectile dysfunction or sexual dysfunction, treating wounds, or causing or promoting hair growth. For example, in one aspect, the present invention is generally directed to compositions for delivery of nitric oxide, transdermally and/or to a mucosal surface. The composition may include nitric oxide. The nitric oxide may be present within a first phase comprising a lecithin, such as phosphatidylcholine. In certain embodiments, the lecithin is present in liposomes, micelles, or other vesicles containing nitric oxide. The composition can take the form of a gel, a cream, a lotion, an ointment, a solution, a solid “stick,” etc., that can be rubbed or sprayed onto the skin, e.g., onto the penis, vulva, or other suitable portion of the skin. The composition can also be applied to a mucosal surface in some instances. In some embodiments, a composition comprising nitric oxide is unexpectedly stable and can be stored long-term. Thus, for example, a composition of the invention may exhibit a long shelf life, with little loss or reaction of nitric oxide. This may be particularly useful for certain applications such as consumer products, including those described herein. Other aspects of the present invention are generally directed to methods of making or using such compositions, methods of promoting such compositions, kits including such compositions, or the like.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 61/453,836, filed Mar. 17, 2011, entitled “TopicalNitric Oxide Systems,” by Nicholas V. Perricone, incorporated herein byreference.

FIELD OF INVENTION

The present invention generally relates to compositions and methods fortransdermal drug delivery.

BACKGROUND

Topical drug delivery systems are known. These systems deliver drugs orother therapeutic agents, or other desired substances transdermally, andmay be designed to act locally at the point of application, or to actsystemically once entering the body's blood circulation. In thesesystems, delivery may be achieved by techniques such as direct topicalapplication of a substance or drug in the form of an ointment or thelike, or by adhesion of a patch with a reservoir or the like that holdsthe drug (or other substance) and releases it to the skin in atime-controlled fashion.

Topical delivery systems for substances such as drugs, pain relievingcompounds, vitamins, and skin improving compounds have been in use for anumber of years. Transdermal delivery systems using creams have beendeveloped for use with analgesics and skin refining compounds. However,these delivery systems do not work effectively for all compounds.

SUMMARY

The present invention generally relates to compositions and methods fortransdermal drug delivery. The compositions can be used in a variety ofapplications. For example, in some embodiments, transdermal systems andmethods are provided for use in treating erectile dysfunction or sexualdysfunction. In other embodiments, transdermal systems and methods areprovided for use in treating wounds. In still other embodiments,transdermal systems and methods are provided for use in causing orpromoting hair growth. The subject matter of the present inventioninvolves, in some cases, interrelated products, alternative solutions toa particular problem, and/or a plurality of different uses of one ormore systems and/or articles.

In some embodiments, aspects of the invention relate to methods andcompositions for delivering nitric oxide transdermally. In one aspect,the present invention is generally directed to compositions fortransdermal delivery that include a lecithin such as phosphatidylcholinethat can contain or “trap” nitric oxide. The phosphatidylcholine orlecithin may be contained within a second phase that causes thephosphatidylcholine or lecithin to form vesicles, e.g., micelles orliposomes. Compositions such as these can be used for variousapplications, for example, to treat sexual dysfunction. For example, inone embodiment, a composition can be applied to the genital region,e.g., perineal and/or other genital region of a subject.

The present invention relates, in another aspect, to a composition fortransdermal delivery of nitric oxide that comprises a lecithin (e.g.,phosphatidylcholine) component entrapping nitric oxide, wherein thelecithin component stabilizes the nitric oxide at room temperature, atrefrigerator temperature (e.g., 4° C.), or at or lower than 80° C., forextended period of storage. The composition may further comprise one ormore adjunct ingredients.

In another aspect, the present invention relates to a method fortreating various diseases that comprises applying to skin a compositioncomprising an effective amount of nitric oxide and a phase having alecithin (e.g., phosphatidylcholine) component entrapping the nitricoxide.

In some embodiments, aspects of the invention relate to a method fortreating a sexual disorder or dysfunction, by contacting a genitalregion of a subject with a composition comprising an emulsion having afirst phase comprising nitric oxide and lecithin, and a second phasecomprising an emulsifier, wherein the lecithin is present at at leastabout 0.25% by weight of the composition, and wherein the first phasecomprises no more than about 250 ppm of water by weight of thecomposition.

In some embodiments, the genital region is a perineal region. In someembodiments, the genital region is a penile surface region. In someembodiments, the genital region is a vaginal surface region.

In some embodiments, the first phase forms discrete vesicles containedwithin the second phase. For example, the first phase can form liposomescontained within the second phase. In some embodiments, the first phaseforms multilamellar liposomes contained within the second phase. In someembodiments, the first phase forms a liquid crystal.

In some embodiments, the first phase comprises no more than about 100ppm of water by weight of the composition.

In some embodiments, the nitric oxide is present at at least about 0.5%by weight of the composition. In other cases, the nitric oxide may bepresent at a concentration of between about 400 mg/kg and about 1000mg/kg, and in some cases, between about 500 mg/kg and about 800 mg/kg.

In some embodiments, at least some of the nitric oxide is present withinthe first phase as a gas. In some embodiments, at least some of thenitric oxide is present within the first phase bound to lecithin. Insome embodiments, at least some of the nitric oxide is present withinthe first phase bound to phosphatidylcholine.

In some embodiments, the lecithin is present at at least about 0.5% byweight. In some embodiments, the lecithin is present at at least about1% by weight. In some embodiments, the lecithin is present at at leastabout 30% by weight. In some embodiments, the lecithin is present at atleast about 60% by weight.

In some embodiments, the lecithin comprises a phosphatidylcholine. Insome embodiments, at least some of the phosphatidylcholine comprises aphosphatidylcholine comprising a stearic diglyceride linked to a cholineester of a phosphoric acid. In some embodiments, at least some of thephosphatidylcholine comprises a phosphatidylcholine comprising apalmitic diglyceride linked to a choline ester of a phosphoric acid. Insome embodiments, at least some of the phosphatidylcholine comprises aphosphatidylcholine comprising an oleic diglyceride linked to a cholineester of a phosphoric acid. In some embodiments, at least some of thephosphatidylcholine comprises a polyenylphosphatidylcholine. In someembodiments, the polyenylphosphatidylcholine comprises linoleic acid. Insome embodiments, the polyenylphosphatidylcholine comprisesdilinoleoylphosphatidylcholine. In some embodiments, at least about 30wt % of the phosphatidylcholine is a polyenylphosphatidylcholine.

In some embodiments, the first phase further comprises a transdermalpenetration enhancer. In some embodiments, the transdermal penetrationenhancer is present at least about 5% by weight of the composition.

In some embodiments, the first phase comprises a fatty acid ester. Insome embodiments, the first phase comprises ascorbate palmitate. In someembodiments, the first phase comprises isopropyl palmitate. In someembodiments, the first phase further comprises1,3-dimethyl-2-imidazolidinone.

In some embodiments, the second phase further comprises 1,2-propanediol.In some embodiments, the second phase further comprises a polyglycol. Insome embodiments, the polyglycol comprises polyethylene glycol.

In some embodiments, the composition is a gel. In some embodiments, thecomposition is a cream. In some embodiments, the composition issubstantially transparent.

In some embodiments, the composition has a viscosity of at least about20,000 cP. In some embodiments, the composition has a viscosity of atleast about 50,000 cP.

In yet another aspect, the present invention encompasses methods ofmaking one or more of the embodiments described herein.

In some embodiments, a lipid composition can be loaded with nitric oxidby passing nitric oxide gas through the lipid composition underconditions wherein the lipid is in a fluid state. In some embodiments,nitric oxide gas is bubbled through a lipid that is in a fluid state. Insome embodiments, nitric oxide gas is bubbled for about 5 hours or less,for example about 4-5 hours, about 3-4 hours, about 2-3 hours, about 1-2hours, about 30-60 minutes, about 15-30 minutes, or about 5-15 minutes.

In another aspect, the present invention is generally directed to amethod for treating diseases, e.g., by applying to skin a compositioncomprising an effective amount of nitric oxide and a carrier having aphosphatidylcholine component entrapping the nitric oxide.

In still another aspect, the present invention is generally directed toa method comprising contacting a genital region of a subject with acomposition comprising an emulsion comprising a first phase comprisingnitric oxide and lecithin, and a second phase comprising an emulsifier.In some cases, the lecithin is present at at least about 0.25% by weightof the composition. In certain embodiments, the first phase comprises nomore than about 250 ppm of water by weight of the composition.

The present invention, according to yet another aspect, is generallydirected to a method comprising contacting a wound on the skin of asubject with a composition comprising an emulsion comprising a firstphase comprising nitric oxide and lecithin, and a second phasecomprising an emulsifier. In some instances, the lecithin is present atat least about 0.25% by weight of the composition. In certainembodiments, the first phase comprises no more than about 250 ppm ofwater by weight of the composition.

According to still another aspect, the present invention is generallydirected to a method comprising contacting the skin of a subject, at alocation where hair growth is desired, with a composition comprising anemulsion comprising a first phase comprising nitric oxide and lecithin,and a second phase comprising an emulsifier. In accordance with certainembodiments, the lecithin is present at at least about 0.25% by weightof the composition. In some cases, the first phase comprises no morethan about 250 ppm of water by weight of the composition.

Yet another aspect of the present invention is generally directed to amethod of formulating a topical nitric oxide composition. For example,in some embodiments, the method comprises acts of mixing a polyglycoland a phosphatidylcholine to form a phosphatidylcholine solution, anddelivering nitric oxide into the phosphatidylcholine solution.

Another aspect of the present invention is generally directed to acomposition for transdermal delivery of nitric oxide. In one set ofembodiments, the composition comprises a phosphatidylcholine carrierentrapping nitric oxide. In some cases, the phosphatidylcholine carrierstabilizes the nitric oxide at a temperature at or lower than 80° C.

In another set of embodiments, the composition comprises an emulsioncomprising a first phase comprising nitric oxide and lecithin, and asecond phase comprising an emulsifier. In some cases, the lecithin ispresent at at least about 0.25% by weight of the composition. In certainembodiments, the first phase comprises no more than about 250 ppm ofwater by weight of the composition.

According to another set of embodiments, the composition comprises anemulsion comprising a first phase comprising a gas and lecithin, and asecond phase comprising an emulsifier. The lecithin may be present at atleast about 0.25% by weight of the composition, in some instances. Invarious embodiments, the gas is present at at least about 0.5% by weightof the composition. In addition, the first phase comprises no more thanabout 250 ppm of water by weight of the composition in some cases.

The composition, in yet another set of embodiments, comprises a firstphase contained within a second phase comprising a fatty acid ester. Incertain cases, the first phase comprises nitric oxide andphosphatidylcholine at at least about 0.25% by weight of thecomposition.

Still another set of embodiments is generally directed to a compositioncomprising a first phase and a second phase substantially immiscible inthe first phase. The first phase may comprise nitric oxide andphosphatidylcholine in certain embodiments.

In another set of embodiments, the composition is generally directed toa gel or a cream comprising nitric oxide. In some instances, the gelcomprises no more than about 250 ppm of water. In some embodiments, thefirst and second phases of a composition each comprise no more thanabout 250 ppm of water. In some embodiments, one or more of thematerials used to prepare a nitric oxide containing composition eachcomprise no more than about 250 ppm of water. In some embodiments, anitric oxide containing composition prepared as described hereincomprises no more than 250 ppm of water.

In still another aspect, the present invention encompasses methods ofusing one or more of the embodiments described herein.

Other advantages and novel features of the present invention will becomeapparent from the following detailed description of various non-limitingembodiments of the invention when considered in conjunction with theaccompanying figures. In cases where the present specification and adocument incorporated by reference include conflicting and/orinconsistent disclosure, the present specification shall control. If twoor more documents incorporated by reference include conflicting and/orinconsistent disclosure with respect to each other, then the documenthaving the later effective date shall control.

DETAILED DESCRIPTION

The present invention generally relates to compositions and methods fortransdermal drug delivery. The compositions can be used in a variety ofapplications, including treating erectile dysfunction or sexualdysfunction, treating wounds, or causing or promoting hair growth. Forexample, in one aspect, the present invention is generally directed tocompositions for delivery of nitric oxide, transdermally and/or to amucosal surface. The composition may include nitric oxide. The nitricoxide may be present within a first phase comprising a lecithin, such asphosphatidylcholine. In certain embodiments, the lecithin is present inliposomes, micelles, or other vesicles containing nitric oxide. Thecomposition can take the form of a gel, a cream, a lotion, an ointment,a solution, a solid “stick,” etc., that can be rubbed or sprayed ontothe skin, e.g., onto the penis, vulva, or other suitable portion of theskin. The composition can also be applied to a mucosal surface in someinstances. In some embodiments, a composition comprising nitric oxide isunexpectedly stable and can be stored long-term. Thus, for example, acomposition of the invention may exhibit a long shelf life, with littleloss or reaction of nitric oxide. This may be particularly useful forcertain applications such as consumer products, including thosedescribed herein. Other aspects of the present invention are generallydirected to methods of making or using such compositions, methods ofpromoting such compositions, kits including such compositions, or thelike.

Unlike prior art that employs only nitric oxide donors as nitric oxidesources, in certain embodiments of the present invention, nitric oxidegas itself is entrapped or contained within various compositions asdiscussed herein, for example, in liquid crystal multilamellarphosphatidylcholine. In addition, in certain embodiments as discussedbelow, the composition may be stable and can be stored for periods oftime with little or no loss or reaction of the nitric oxide containedtherein.

In one aspect, the present invention is useful for treating erectiledysfunction or sexual dysfunction. For example, the composition can beapplied to the skin of a subject on or near a male or female genitalregion to treat sexual dysfunction and/or to enhance sexual performanceor experience. In some embodiments, a composition is applied to theperineal region (e.g., penis), to the vulva, etc., to release nitricoxide to the skin and underlying tissue directly upon topicaladministration of the composition to the subject. For example, acomposition of the invention may be applied to the genital perinealregion or penis or portion thereof of a male subject to treat erectiledysfunction, to the vulva of a female subject to treat sexualdysfunction (or to any other portion at or near the genital perinealregion in male or female subjects).

The composition, in yet another aspect, can be applied to the skin of asubject, for example, to a wound to promote wound healing. In someembodiments, compositions of the invention can be used to promote anywound healing by applying the composition to an existing wound. Itshould be appreciated that compositions of the invention may be used totreat or promote healing of any type of wound, including cuts, scrapes,other traumatic wounds, burns or other accidental wounds. Non-limitingexamples of wounds include wound can be an anal fissure, a surgicalsite, a trauma site, a burn, an abrasion, a sunburn, a cut or lacerationon the skin, or any other damaged region of the skin. In someembodiments, the composition is applied to a mucosal surface of thesubject, for example, to the nose. Accordingly, compositions of theinvention also may be used to treat wounds that result from surgicalintervention (e.g., any medical intervention or operation that requireswound healing as part of the recovery process). It should be appreciatedthat compositions of the invention can be used prophylactically to asite (for example a surgical site) in anticipation of a wound.Accordingly, in some embodiments, a composition of the invention can beused to prepare a tissue site for wound healing prior to the wound(e.g., prior to surgery) and/or after the wound (e.g., and/or aftersurgery). In some embodiments, a composition of the invention may beapplied to the surface of a wound or to skin prior to a wound.

In some embodiments, a composition of the invention may be applied to asurgical device, tool, or other substrate. For example, a composition ofthe invention may be applied to sutures, implants, surgical tools, orother substrates that may come into contact with wounded tissue (e.g.,cut tissue) during surgery. In some embodiments, compositions of theinvention may be provided as a cream or ointment as described in moredetail herein. It also should be appreciated that compositions of theinvention may be provided on surgical dressings, bandages, or othermaterial that is to be contacted to a surgical wound. In someembodiments, a composition may be applied to a material or substrateimmediately prior to use on a subject. However, in some embodiments, amaterial or substrate may be prepared (e.g., packaged, stored, orotherwise prepared) to contain a composition of the invention prior touse. For example, prepackaged bandages or surgical devices, sutures, orimplants may be prepared and packaged with a coating of a composition ofthe invention. Compositions of the invention may be used for human orother animal subjects (male or female).

In still another aspect, a composition of the present invention may beapplied to a subject to promote hair growth, for example in a regionwhere hair growth is desired. In some embodiments, a composition can beapplied to the skin of a subject, for example, to prevent or reduce hairloss, for example in a region where hair loss is undesirable. In someembodiments, a region targeted for hair growth and or a reduction inhair loss may be the skin surface of the scalp. In some embodiments, asubject may be a human. The subject may be male or female. In someembodiments, a subject may have lost hair (e.g., is bald, balding, orhas thinning hair) and/or may be at risk of losing hair (e.g., due toage, a genetic risk and/or family history of hair loss). In someembodiments, the subject may have lost hair or be at risk for losinghair due to injury (e.g., trauma or burn) or due to a disease and/ortreatment (e.g., chemotherapy).

According to some aspects of the invention, nitric oxide is useful as atherapeutic agent useful for in the promotion of healthy cells andtissues and/or because nitric oxide can act to increase local bloodflow. It is generally believed that the therapeutic effect of nitricoxide owes largely to its cell signaling function, although there mayalso be other modes of action. Because nitric oxide is highly reactive(having a lifetime of a few seconds), yet diffuses freely across cellmembranes, nitric oxide often acts as a transient paracrine (betweenadjacent cells) or autocrine (within a single cell) signaling molecule.Nitric oxide produced by one cell can penetrate through cell membranesto regulate the function of other cells, which may lead to cascadingeffects in biological systems. These are also known as the so-callednitric oxide pathways. For example, the endothelium (inner lining) ofblood vessels can use nitric oxide to signal the surrounding smoothmuscle to relax, thus resulting in vasodilation and increasing bloodflow. Nitric oxide also may contribute to vessel homeostasis byinhibiting vascular smooth muscle contraction and growth, plateletaggregation, and leukocyte adhesion to the endothelium.

The human body produces nitric oxide naturally as needed. Nitric oxidemay be biosynthesized endogenously from L-arginine, oxygen, and/or NADPHby various nitric oxide synthetase enzymes. Nitric oxide may also begenerated by phagocytes (monocytes, macrophages, and neutrophils) aspart of the human immune response. However, there often exists a need toprovide additional quantities of nitric oxide for therapeutic reasons,or as dietary supplements. For example, nitric oxide may be introducedto subjects to stimulate or support vasodilatation, anti-inflammation,anti-bacteria, neurotransmission, modulation of the hair cycle,treatment of anal disorders and penile erections (through nitric oxide'sability to vasodilate), or the like. As dietary supplements, nitricoxide training gels, such as FORCE FACTOR® sold by General NutritionCenters (GNC), are taken by athletes and others in the belief thatnitric oxide will increase muscle strength and endurance, reduce warm-uprelated injuries, or decrease recuperation time.

Because nitric oxide is an unstable and reactive gas, therapeuticdelivery of nitric oxide is usually accomplished by dosing a nitricoxide prodrug, also known as a nitric oxide donor, although in somecases, nitric oxide itself can be administrated by inhalation. Thenitric oxide donor is typically formulated in a pharmaceuticallyaccepted carrier for oral, topical, or other administration routes, andthe nitric oxide donor subsequently releases nitric oxide afteradministration. Examples of such nitric oxide donors include L-arginine,nitroglycerin, and amyl nitrite. For subjects that suffer from symptomsthat appear on skin (e.g. anal fissure) or the scalp (e.g. baldness),direct delivery of nitric oxide to the affected skin or scalp can bemore effective and convenient, especially if long-term treatment isrequired. Topical application of nitric oxide can also reduce oreliminate the side effects associated with sustained increased levels ofnitric oxide in the human body. For example, it has been discovered thathigh levels of nitric oxide may result in direct tissue damage and/orcontribute to the vascular collapse associated with septic shock.

Until now, the art has been deficient in topical nitric oxidetreatments. Some aspects of the present invention address this gap inthe art and the therapeutic need by providing various compositions andmethods that deliver nitric oxide topically. In some cases, thesecompositions and methods stimulate the absorption of nitric oxide intocells and tissues, and/or promote vasodilation of skin tissues. Certainaspects of the present invention thus permit treatment or prevention ofvarious symptoms such as erectile dysfunction, hair loss, or woundhealing.

Since nitric oxide is an unstable and reactive gas, entrapment, storage,and release of nitric oxide requires careful formulation in someembodiments of the invention. For example, nitric oxide readily reactswith water to form nitrous acid (HNO₂), and thus, certain embodiments ofthe invention include compositions or phases that are substantially freeof water. As another example, in one set of embodiments, nitric oxidemay be contained within a first phase comprising a lecithin such asphosphatidylcholine, which may be present within a second phasecomprising an emulsifier, such as is discussed herein. Other components,for example, transdermal penetration enhancers, adjuvants, surfactants,lubricants, etc. can also be present in certain cases.

Thus, the compositions of the invention comprise, in certain aspects, aphase comprising phosphatidylcholine and/or other lecithins in whichnitric oxide is contained within or “trapped.” The phosphatidylcholineor lecithin may be contained within a second phase, for example,comprising an emulsifier, which may cause the phosphatidylcholine orlecithin to form vesicles, e.g., micelles or liposomes. Thephosphatidylcholine or lecithin composition can be unilamellar ormultilamellar in some embodiments. In some instances, the presence ofthe second phase causes the phosphatidylcholine or lecithin to form aliquid crystal arrangement.

The nitric oxide is typically gaseous, and may be present within thecomposition as small bubbles and/or bound to lecithins orphosphatidylcholines within the composition. For example, the nitricoxide may be bound to double bonds present in the lecithins orphosphatidylcholines. Phosphatidylcholine is believed to stabilizeand/or contain the nitric oxide. In some cases, stability of thecomposition can be achieved at room temperature (about 25° C.), and/orat other temperatures such as those described herein. Without wishing tobe bound by any theory, it is believed that the phosphatidylcholineadopts a liquid crystal structure under such conditions, which canthereby contain the nitric oxide, e.g., as small gaseous bubbles, and/orthrough binding with lecithins or phosphatidylcholines.

Nitric oxide is typically reactive with water (e.g., forming nitrousacid), which contributes to its relatively short lifetime within thebody or within other aqueous environments. Accordingly, in certainembodiments of the invention, the composition, or at least a phase ofthe composition comprising the nitric oxide (and/or the second phase,and/or one or more materials used to prepare a nitric oxide composition,and/or a nitric oxide composition prepared as described herein), issubstantially free of water, e.g., comprising no more than about 10 wt%, no more than about 3 wt %, no more than about 1 wt %, no more thanabout 0.3 wt %, or no more than about 0.1 wt % water (i.e., relative tothe weight of the overall composition). The composition may also have nomore than about 1,000 ppm, no more than about 750 ppm, no more thanabout 500 ppm, no more than about 400 ppm, no more than about 300 ppm,no more than about 250 ppm, no more than about 200 ppm, no more thanabout 150 ppm, no more than about 100 ppm, no more than about 50 ppm, nomore than about 25 ppm, or no more than about 10 ppm of water. Incertain embodiments, no detectable water may be present in thecomposition, or at least within a phase of the composition comprisingthe nitric oxide. Any suitable technique can be used for determining theamount of water present in the composition, for example, Karl-Fishertitration. In some cases, the composition may also be free of anyliquids that typically contain water, e.g., physiological buffers, bodyfluids, saline, or the like.

Any suitable amount of nitric oxide may be present within a compositionprepared as described herein. For example, at least about 0.3 wt %, atleast about 0.5 wt %, at least about 0.7 wt %, at least about 1 wt %, atleast about 1.5 wt %, at least about 2 wt %, at least about 2.5 wt %, atleast about 3 wt %, at least about 5 wt % at least about 10 wt %, atleast about 20 wt %, at least about 30 wt %, at least about 40 wt %, atleast about 50 wt %, at least about 60 wt %, at least about 70 wt %, atleast about 80 wt %, at least about 90 wt %, at least about 100 wt %, atleast about 110 wt %, or at least about 120 wt % of the composition canbe nitric oxide, where the basis of the weight percentage is the weightof the composition before nitric oxide is added. For example, the nitricoxide may be present at between 70 wt % and about 120 wt % of thecomposition. In some embodiments, the nitric oxide may be present at aconcentration of at least about 400 mg/kg, at least about 450 mg/kg, atleast about 500 mg/kg, at least about 550 mg/kg, at least about 570mg/kg, at least about 600 mg/kg, at least about 650 mg/kg, at leastabout 700 mg/kg, at least about 750 mg/kg, at least about 800 mg/kg, atleast about 850 mg/kg, at least about 950 mg/kg, or at least about 1000mg/kg of the composition. In certain cases, the nitric oxide may bepresent at a concentration of no more than about 2000 mg/kg, no morethan about 1500 mg/kg, no more than about 1000 mg/kg, no more than about960 mg/kg, no more than about 900 mg/kg, no more than about 800 mg/kg,no more than about 700 mg/kg, or no more than about 600 mg/kg. Forexample, the nitric oxide may be present at a concentration of betweenabout 570 mg/kg and about 960 mg/kg.

In some embodiments, nitric oxide is present within a first phasecomprising a lecithin, such as phosphatidylcholine. Phosphatidylcholine(herein abbreviated “PC”) is a basic component of cell membrane bilayersand the main phospholipid circulating in the plasma of blood.Phosphatidylcholine typically has a phospholipid structure with acholine head group and a glycerophosphoric acid tail group. The tailgroup can be saturated or unsaturated. More than one tail group may bepresent in the phosphatidylcholine in some cases, and the tail groupsmay be the same or different. Specific non-limiting examples ofphosphatidylcholines that could be used include one or a mixture ofstearic, palmitic, margaric, and/or oleic acid diglycerides linked to acholine ester head group.

Phosphatidylcholines are a member of a class of compounds calledlecithins. Typically, a lecithin is a composed of phosphoric acid,choline, fatty acids, glycerol, glycolipids, triglycerides, and/orphospholipids. In some cases, other lecithins may be used, in additionto or instead of a phosphatidylcholine. Non-limiting examples of otherlecithins include phosphatidylethanolamine, phosphatidylinositol, orphosphatidic acid. Many commercial lecithin products are available, suchas, for example, Lecithol®, Vitellin®, Kelecin®, and Granulestin®.Lecithin is widely used in the food industry. In some embodiments,certain compositions of the invention can contain synthetic or naturallecithin, or mixtures thereof. Natural preparations are used in somecases because they exhibit desirable physical characteristics, and/ormay be economical or nontoxic. However, in other embodiments,non-natural preparations are used, or the composition can include bothnatural and non-natural preparations.

Any suitable amount of phosphatidylcholine or lecithin may be presentwithin the composition. For example, at least about 0.25 wt %, at leastabout 0.5 wt %, at least about 1 wt %, at least about 2 wt %, at leastabout 3 wt %, at least about 5 wt %, at least about 8 wt %, at leastabout 10 wt %, at least about 20 wt %, at least about 30 wt %, at leastabout 40 wt %, at least about 50 wt %, at least about 60 wt %, at leastabout 70 wt %, at least about 80 wt %, or at least about 90 wt % of theentire composition can be a phosphatidylcholine or a lecithin. In somecases, the phosphatidylcholine or lecithin may be present at aconcentration of no more than about 95 wt %, no more than about 90 wt %,no more than about 80 wt %, no more than about 70 wt %, no more thanabout 65 wt %, no more than about 60 wt %, no more than about 50 wt %,no more than about 40 wt %, no more than about 30 wt %, no more thanabout 20 wt %, or no more than about 10%. For instance, thephosphatidylcholine or lecithin may be present at between about 8 wt %and about 65 wt %, or between about 0 wt % and about 10 wt %, etc. Oneor more than one type of phosphatidylcholine or lecithin may be present.

Some delivery compositions of the present invention may containpolyenylphosphatidylcholine (herein abbreviated “PPC”). In some cases,PPC can be used to enhance epidermal penetration. The term“polyenylphosphatidylcholine,” as used herein, means anyphosphatidylcholine bearing two fatty acid moieties, wherein at leastone of the two fatty acids is an unsaturated fatty acid with at leasttwo double bonds in its structure, such as linoleic acid.

Certain types of soybean lecithin and soybean fractions, for example,can contain higher levels of polyenylphosphatidylcholine, withdilinoleoylphosphatidylcholine (18:2-18:2 phosphatidylcholine) as themost abundant phosphatidylcholine species therein, than conventionalfood grade lecithin. Such lecithins may be useful in formulating certaindelivery compositions. In some embodiments, conventional soybeanlecithin may be enriched with polyenylphosphatidylcholine, for instance,by adding soybean extracts containing high levels ofpolyenylphosphatidylcholine. As used herein, this type ofphosphatidylcholine is called “polyenylphosphatidylcholine-enriched”phosphatidylcholine (hereinafter referred to as PPC-enrichedphosphatidylcholine), even where the term encompasses lecithin obtainedfrom natural sources exhibiting polyenylphosphatidylcholine levelshigher than ordinary soybean varieties. These products are commerciallyavailable, for example, from American Lecithin Company, Rhone-Poulencand other lecithin vendors. American Lecithin Company markets itsproducts with a “U” designation, indicating high levels of unsaturation;Rhone-Poulenc's product is a soybean extract containing about 42%dilinoleoylphosphatidylcholine and about 24%palmitoyllinoleylphosphatidylcholine (16:0 to 18:2 of PC) as the majorphosphatidylcholine components. Another example of a suitablepolyenylphosphatidylcholine is NAT 8729 (also commercially availablefrom vendors such as Rhone-Poulenc and American Lecithin Company).

Any suitable amount of polyenylphosphatidylcholine may be present withinthe composition. For example, at least about 0.25 wt %, at least about0.5 wt %, at least about 1 wt %, at least about 2 wt %, at least about 3wt %, at least about 5 wt %, at least about 8 wt %, at least about 10 wt%, at least about 20 wt %, at least about 30 wt %, at least about 40 wt%, at least about 50 wt %, at least about 60 wt %, at least about 70 wt%, at least about 80 wt %, or at least about 90 wt % of the compositioncan be polyenylphosphatidylcholine. In some cases, thepolyenylphosphatidylcholine may be present at a concentration of no morethan about 95 wt %, no more than about 90 wt %, no more than about 80 wt%, no more than about 70 wt %, no more than about 65 wt %, no more thanabout 60 wt %, no more than about 50 wt %, no more than about 40 wt %,no more than about 30 wt %, no more than about 20 wt %, or no more thanabout 10%. For instance, the polyenylphosphatidylcholine may be presentat between about 8 wt % and about 65 wt %. In some embodiments, at leastabout 20 wt %, at least about 30 wt %, at least about 40 wt %, at leastabout 50 wt %, at least about 60 wt %, at least about 70 wt %, at leastabout 80 wt %, at least about 90 wt %, or about 100 wt % of all of thephosphatidylcholine or lecithin in the composition ispolyenylphosphatidylcholine.

While not wishing to be bound to any theory, it is believed that thePPC-enriched phosphatidylcholine forms a bilayer enveloping nitric oxide(and in some embodiments, other adjunct ingredients, if present) tocreate the drug delivery composition. The PPC-enrichedphosphatidylcholine is believed to contribute to the stability of thenitric oxide, for example, by shielding the nitric oxide from water,and/or by enhancing its penetration into the skin or other area, e.g., amucosal surface.

The first phase also comprises, in some embodiments of the invention, afatty acid ester. Non-limiting examples include ascorbate palmitate orisopropyl palmitate. In some cases, the fatty acid ester is used as apreservative or an antioxidant. The composition can include any suitableamount of fatty acid ester, for example, at least about 1 wt %, at leastabout 3 wt %, at least about 5 wt %, at least about 10 wt %, at leastabout 20 wt %, at least about 30 wt %, at least about 40 wt %, at leastabout 50 wt %, etc. In some cases, no more than about 60 wt %, no morethan about 50 wt %, no more than about 40 wt %, no more than about 30 wt%, no more than about 20 wt %, no more than about 18 wt %, no more thanabout 15 wt %, no more than about 12 wt %, or no more than about 10 wt %of the composition is fatty acid ester. For example, the composition maybe between about 0 wt % and about 10 wt % fatty acid ester. Thecomposition may include one or more than one fatty acid ester.

In certain embodiments, a drug delivery composition such as thosedescribed herein can be formulated to include a second phase. Typically,the second phase is substantially immiscible with the first phasecomprising phosphatidylcholine or lecithin. Two phases that aresubstantially immiscible are able to form discrete phases when exposedto each other at ambient conditions (e.g., 25° C. and 1 atm) forextended periods of time (e.g., at least about a day). The phases can beseparate identifiable phases (e.g., one may float above the other), orin some cases, the phases are intermingled, e.g., as in an emulsion. Thestability of the discrete phases may be kinetic and/or thermodynamic innature, in various embodiments.

In one set of embodiments, the second phase may comprise an emulsifierwhich causes the first phase comprising phosphatidylcholine or lecithinto form a liquid crystal, and/or vesicles such as micelles or liposomes.Typically, in a liquid crystal phase, vesicular structures such asmicelles, liposomes, hexagonal phases, or lipid bilayers can be formed.In some cases, multilamellar structures may be present within the liquidcrystal phase, although in other cases, only unilamellar structures maybe present. For example, in certain cases, the PPC-enrichedphosphatidylcholine can be loosely arranged in a multilamellar fashion,with nitric oxide and optional adjunct ingredients being bonded orotherwise entrapped or contained within the lipid bilayers formedtherein. In some cases, the first phase (e.g., comprising PPC-enrichedphosphatidylcholine) and the second phase can form a structure such asis disclosed in U.S. Pat. No. 7,182,956 to Perricone, et al. This isbelieved (without wishing to be bound by any theory) to form a looselyarranged, yet stable, PPC-enriched phosphatidylcholine-drug complex thatmay allow penetration and delivery of nitric oxide and optional adjunctingredients to the skin, e.g., to the dermal vasculature, or to amucosal surface.

In one set of embodiments, the second phase comprises an emulsifier. Theemulsifier, in one embodiment, is a substance that is able to stabilizean emulsion by increasing its kinetic stability. The emulsifier may alsobe chosen in some cases to be relatively inert or non-toxic relative tothe skin or to a mucosal surface.

In some embodiments, the second phase may comprise a polyglycol. Thepolyglycol may include a polyhydric alcohol of a monomeric glycol suchas polyethylene glycol (PEG) and/or polypropylene glycol (PPG). Forexample, the PEG or PPG may be PEG or PPG 200, 300, 400, 600, 1,000,1,450, 3,350, 4,000, 6,000, 8,000, and 20,000, where the numberindicates the approximate average molecular weight of the PEG or PPG. Asis understood by those of ordinary skill in the art, a polyglycolcomposition often will comprise a range of molecular weights, althoughthe approximate average molecular weight is used to identify the typepolyglycol. More than one PEG and/or PPG can also be present in certaininstances.

The second phase may comprise a surfactant in some embodiments.Non-limiting examples of surfactants include a siloxylated polyethercomprising dimethyl, methyl(propylpolyethylene oxide propylene oxide,acetate) siloxane commercially available from vendors such as DowCorning (Dow Corning 190 surfactant). Other examples of materials thatcan be used as (or within) the second phase include, but are not limitedto, 1,2-propanediol, or silicone fluids containing low viscositypolydimethylsiloxane polymers, methylparaben (p-hydroxy benzoic acidmethyl ester) commercially available from vendors such as Dow Corning(Dow Corning 200 silicone fluid). Still other examples include varioussiloxane or silicone compounds, e.g., hexamethyldisiloxane,amodimethicone, phenyltrimethicone, etc.

Additionally, purified water may be added to the second phase in someembodiments, although in other cases, little or no water is present inthe second phase. For example, the first phase, the second phase, cancontain less than 10%, less than 5%, less than 2%, less than 1%, or lessthat 0.05% (e.g., wt %) of water relative to the weight of therespective phase or of the entire composition. In some cases, the secondphase may also comprise adjunct ingredients such as those describedherein.

The second phase may include any one, or more than one, of the materialsdescribed above. In addition, any suitable amount of second phase can beused in accordance with various embodiments of the invention. Forexample, the second phase may be present at at least about 10 wt %, atleast about 20 wt %, at least about 30 wt %, at least about 40 wt %, atleast about 50 wt %, at least about 60 wt %, at least about 70 wt %, atleast about 80 wt %, or at least about 90 wt % of the composition. Insome cases, the ratio of the first phase (e.g., comprisingphosphatidylcholine or lecithin) to the second phase can be at leastabout 1:3, at least about 1:2, at least about 1:1, at least about 2:1,at least about 3:1, or at least about 4:1, etc.

In another set of embodiments, the composition may also include one ormore transdermal penetration enhancers. Examples of transdermalpenetration enhancers include, but are not limited to,1,3-dimethyl-2-imidazolidinone or 1,2-propanediol. Other examplesinclude cationic, anionic, or nonionic surfactants (e.g., sodium dodecylsulfate, polyoxamers, etc.); fatty acids and alcohols (e.g., ethanol,oleic acid, lauric acid, liposomes, etc.); anticholinergic agents (e.g.,benzilonium bromide, oxyphenonium bromide); alkanones (e.g., n-heptane);amides (e.g., urea, N,N-dimethyl-m-toluamide); organic acids (e.g.,citric acid); sulfoxides (e.g., dimethylsulfoxide); terpenes (e.g.,cyclohexene); ureas; sugars; carbohydrates or other agents. Thetransdermal penetration enhancers can be present in any suitable amountwithin the composition. For example, at least about 10 wt %, at leastabout 20 wt %, at least about 30 wt %, at least about 40 wt %, or atleast about 50 wt % of the composition may comprise one or moretransdermal penetration enhancers. In some cases, no more than about 60wt %, no more than about 50 wt %, no more than about 40 wt %, no morethan about 30 wt %, no more than about 20 wt %, no more than about 10 wt%, no more than about 9 wt %, or no more than about 5 wt % of thecomposition comprises transdermal penetration enhancers. For example,the composition may have between about 0 wt % and about 5 wt % of one ormore transdermal penetration enhancers.

As a specific non-limiting example of one set of embodiments, apolyenylphosphatidylcholine comprises a certain material with the tradename NAT 8729, and optionally at least one polyglycol (polyhydricalcohol of a monomeric glycol such as polyethylene glycol 200, 300, 400,600, 1,000, 1,450, 3,350, 4,000, 6,000, 8,000 and 20,000). Thecomposition can also comprise a PPC-enriched phosphatidylcholinematerial that is present within the first or second phase, e.g.,comprising nitric oxide. The second phase may also comprise a surfactantsuch as a siloxylated polyether comprising dimethyl,methyl(propylpolyethylene oxide propylene oxide, acetate) siloxanecommercially available from vendors such as Dow Corning (Dow Corning 190surfactant) and lubricant such as silicone fluids containing lowviscosity polydimethylsiloxane polymers, methylparaben (p-hydroxybenzoic acid methyl ester) commercially available from vendors such asDown Corning (Dow Corning 200 silicone fluid).

In some embodiments, various compositions of the invention areformulated to be substantially clear or substantially transparent.Transparency may be useful, for instance, for product acceptance in themarketplace, e.g., when applied to the skin of a subject. However, inother embodiments, the composition is not necessarily transparent.Certain substances can be useful in providing a substantiallytransparent composition, for example, fatty acid esters such asascorbate palmitate or isopropyl palmitate. In one set of embodiments,the composition may be substantially transparent such that incidentvisible light (e.g., have wavelengths of between about 400 nm and about700 nm) can be transmitted through 1 cm of the composition with a lossin intensity of no more than about 50%, about 60%, about 70%, about 80%,or about 90% relative to the incident light. In some embodiments, theremay be no substantial difference in the wavelengths that are absorbed bythe composition (i.e., white light passing through the compositionappears white), although in other cases, there can be more absorption atvarious wavelengths (for example, such that white light passing throughthe composition may appear colored).

Other components may also be present within the composition, inaccordance with certain embodiments of the invention. For example, thecomposition may include volatile organic fluids, fatty acids, volatilearomatic cyclic compounds, high molecular weight hydrocarbons, or thelike.

In accordance with certain aspects of the invention, the composition maybe prepared by mixing a first phase and a second phase together, thenpassing nitric oxide through the mixture. As discussed above, the secondphase can comprise an emulsifier, or any other components discussedherein. The first phase may comprise a lecithin such asphosphatidylcholine and/or polyenylphosphatidylcholine, e.g.,PPC-enriched phosphatidylcholine, for instance, as described herein. Insome embodiments, other components are also mixed into the composition,before or after (or while) adding nitric oxide, for example, transdermalpenetration enhancers, adjuvants, polyglycols (e.g., PEG and/or PPG),surfactants, lubricants, etc. as discussed herein. In some embodiments,however, nitric oxide may be passed through the first phase prior tomixing of the first phase with the second phase.

In one set of embodiments, after forming the mixture, nitric oxide canbe passed into or through the mixture, for example, by blowing bubblesof nitric oxide through the mixture. Nitric oxide may be delivered intothe mixture under pressures such as between about 3,000 Pa and about15,000 Pa, between about 5,000 Pa and about 10,000 Pa, or between about6,000 Pa and about 8,000 Pa, and/or temperatures such as between about0° C. and about 50° C., between about 20° C. and about 35° C., or about25° C. and about 30° C. However, higher or lower pressures also may beused in some embodiments as aspects of the invention are not limited inthis respect.

In certain embodiments, the nitric oxide is bubbled through the mixtureuntil the mixture begins to at least partially solidify. As an example,the viscosity of the mixture may increase to at least about 1,000 cP, atleast about 2,000 cP, at least about 3,000 cP, at least about 5,000 cP,at least about 7,000 cP, at least about 10,000 cP, at least about 12,000cP, at least about 15,000 cP, at least about 20,000 cP, at least about30,000 cP, at least about 40,000 cP, at least about 50,000 cP, at leastabout 60,000 cP, at least about 70,000 cP, or at least about 80,000 cP.The nitric oxide can be passed through the mixture as pure nitric oxide,and/or with other gases (e.g., a noble gas, for example, argon). In somecases, a nitric oxide donor may be passed into the mixture, and therein,at least some of the nitric oxide donor can be converted into nitricoxide. In other embodiments, however, the final composition may havelower viscosities, for example, such that the composition is liquid, orcould be sprayed onto the skin or onto a mucosal surface.

In one set of embodiments, the nitric oxide can be bubbled through themixture to cause the viscosity of the mixture to increase. For example,the viscosity can increase until the mixture begins to form a gel, acream, a lotion, an ointment, a solid “stick,” or the like. A cream maybe, for example, a semi-solid emulsion, e.g., comprising a first phaseand a second phase. The first phase may be discontinuous (e.g.,comprising small droplets or vesicles, such as is discussed herein) andthe second phase may be continuous, or vice versa. In some cases,however, both the first phase and the second phase are co-continuouswithin the mixture.

In some embodiments of the invention, a composition may be prepared asdiscussed above, then diluted, e.g., with a diluent, to produce a finalcomposition. For example, a “stock” composition may be initiallyprepared, e.g., having a relatively high nitric oxide concentration,then the stock composition diluted to produce a final composition, e.g.,before use, before storage, before packaging, etc. In some embodiments,the diluent used may be a component as discussed herein (for example,forming at least a portion of the second phase), and the same ordifferent materials than may be present in the initial composition maybe used. The dilution ratio (amount of diluent added, relative to theinitial composition) may be at least about 2, at least about 3, at leastabout 5, at least about 10, at least about 15, at least about 20, atleast about 25, at least about 30, at least about 50, or at least about100, or any other suitable factor.

A composition may be prepared and/or stored at any suitable temperatureand under any suitable conditions. In some embodiments, for instance, acomposition can be prepared and/or stored under limited or no oxygenconditions, as oxygen can adversely react with nitric oxide. Thecomposition can also be prepared and/or stored under limited or nonitrogen and/or carbon dioxide, as both can also react adversely withnitric oxide. For instance, the composition may be prepared and/orstored in a sealed environment (e.g., stored in a sealed container). Thesealed environment (e.g., container) can be at least substantiallydevoid of gas, and/or contains a gaseous mixture that excludes, or atleast is depleted in, oxygen. In some embodiments, an environmentdepleted in oxygen may have less than about 20%, less than about 15%,less than about 10%, less than about 5%, about 1% or less, about 0.1% orless, about 0.01% or less, about 0.001% or less, oxygen (e.g., as a wt %or as molar % per volume). For example, the gaseous mixture may includea noble gas, such as argon, helium, neon, etc. In one set ofembodiments, the container may comprise a multi-layered metallic and/orpolymeric barrier, e.g., formed from Glaminate® (American Can Company).For instance, the container may have the shape of a tube. Thus, incertain embodiments, the container is substantially resistant to oxygenpermeation, nitrogen permeation, and/or carbon dioxide permeation. Incertain embodiments, the container is substantially watertight, forexample, such that substantially no water is absorbed by the container,or such that no water is able to pass through the container even if thecontainer is filled with water.

As previously discussed, nitric oxide can react with water, and thus,compositions described herein may be prepared and/or stored underconditions where substantially no water is present. For example, nitricoxide and/or a nitric oxide containing preparation described herein maybe prepared and/or stored under relatively low relative humidities(e.g., less than about 50% RH, less than about 40% RH, less than about30% RH, less than about 20% RH, or less than about 10% RH), and/or inthe presence of a suitable desiccant, such as phosphorous pentoxide orsilica gel.

In certain embodiments, the mixture may be mixed with or otherwiseinclude adjunct ingredients, if applicable, and nitric oxide may beintroduced to the mixture, e.g., bubbles of nitric oxide gas may beblown into the mixture until the mixture hardens to obtain the desiredfinal composition. As a specific non-limiting example, a nitric oxidecomposition may be formed by preparing a non-liposome multilamellarliquid crystal phosphatidylcholine phase, for example, by providing apolyglycol, then introducing phosphatidyl choline into the glycol atroom temperature to form a phosphatidylcholine solution. Thephosphatidylcholine often comes as a solid (e.g., as a “brick” ofmaterial), and the phosphatidylcholine may be broken down into smallerpieces to aid in mixing, e.g., by “shaving” or grinding thephosphatidylcholine solid. The phosphatidylcholine solution is mixeduntil the phosphatidylcholine solution is substantially clear, then onemay warm the phosphatidylcholine solution to 40° C., mill the warmedsolution (i.e., low agitation after the initial mixing), combinesiloxylated polyether and polydimethylsiloxane to form a fluid, add thefluid to the warmed solution and milling until the solution is clear,adding methyl paraben or other suitable lubricant to the solution andmilling until the methyl paraben dissolves in the solution, warm waterto 40° C. and adding the warmed water slowly to the solution, and thenceasing milling of the solution and “sweeping” the solution (e.g., witha sweep mixer) to cool to room temperature. Nitric oxide gas can then bebubbled or otherwise introduced into the solution while cooling thesolution until the solution begins to harden or becomes stiff, e.g.,having the consistency of a gel or a cream, such as previouslydescribed. In some cases, the resulting composition is sealed in acontainer, for example, as discussed herein. Any suitable container maybe used, e.g., a tube or a bottle. In addition, the composition (e.g.,within the container) may be stored at room temperature, or any othersuitable temperature. For example, a composition of the invention may bestored at or below 80° C., e.g., at or below room temperature (about 25°C.) or in a refrigerator (e.g., at 4° C.) for extended period ofstorage, for instance, to prevent nitric oxide leakage or denaturing.

It is surprising that, according to some embodiments, nitric oxide notonly can be entrapped in phosphatidylcholine or lecithin compositionssuch as those described herein, but also that such entrappedcompositions may have a long shelf life, especially when refrigerated.No loss or reaction of nitric oxide is expected during extendedrefrigerated storage, at least under certain conditions. For instance,in certain embodiments, the composition may be stored at temperatures ofless than about 80° C., less than about 70° C., less than about 60° C.,less than about 50° C., less than about 40° C., less than about 30° C.,less than about 25° C., less than about 20° C., less than about 15° C.,less than about 10° C., less than about 5° C., less than about 0° C.,etc., for extended periods of time, e.g., at least about a day, at leastabout a week, at least about 4 weeks, at least about 6 months, etc.

Without wishing to be bound by theory, it is believed that nitric oxideforms reversible physical bonds, similar to hydrogen bonds or van derWaals forces, with phosphatidylcholine or other lecithin molecules,e.g., containing one or more double bonds, which may allow nitric oxideto become entrapped and thereby remain intact for an extended period oftime, e.g., during storage. These physical bonds, however, are believedto be not very stable, and may in some cases be easily broken up, forexample, upon various physical agitations such as rubbing thecomposition against skin or a mucosal surface, thereby releasing theentrapped nitric oxide. While others have stabilized other substances ordrugs within phosphatidylcholine or lecithin compositions or vesicles,for example, protein drugs such as insulin, it is surprising that asmall, highly reactive molecule such as NO could similarly bestabilized, especially when it would have been expected that a moleculeas small as NO would readily diffuse away from such compositions and/orwould have reacted with water that is typically present within suchcompositions.

In some embodiments, it is believed that other species reactive withwater could also be similarly stabilized, e.g., within a composition asherein described. Any species that ordinarily reacts with water could bestabilized within such compositions. Examples of such species include,but are not limited to, lithium, or drugs or polymers with labile bondssusceptible to hydrolysis, for instance, certain peptides,polysaccharides, polylactic acid, polyglycolic acid, etc.

In certain aspects of the invention, a composition such as thosedescribed herein can be administered to a subject, such as a humansubject, by rubbing it on the skin or a mucosal surface of the subject,e.g., in areas located at or at least within the vicinity of a desiredtarget area. Without wishing to be bound by any theory, it is believedthat phosphatidylcholine provides or facilitates delivery of nitricoxide to the skin or a mucosal surface, and/or to tissues below the skinor mucosal surface, allowing nitric oxide to be delivered to a targetarea. In some embodiments, the composition can be applied, by rubbingthe composition topically against the skin, or to the mucosal surface,which allows the composition (or at least, nitric oxide) to be absorbedby the skin or mucosal surface. The composition can be applied once, ormore than once. For example, the composition may be administered atpredetermined intervals. In some embodiments, for instance, thecomposition may be applied once per day, twice per day, 3 times per day,4 times per day, once every other day, once every three days, once everyfour days, etc. The amount of nitric oxide necessary to bring about thetherapeutic treatment is not fixed per se, and may depend upon factorssuch as the desired outcome, the type and severity the disease orcondition, the form of nitric oxide, the concentration of nitric oxidepresent within the composition, etc.

Thus, another aspect of the invention provides methods of administeringany composition such as is discussed herein to a subject. Whenadministered, the compositions of the invention are applied in atherapeutically effective, pharmaceutically acceptable amount as apharmaceutically acceptable formulation. Any of the compositions of thepresent invention may be administered to the subject in atherapeutically effective dose. When administered to a subject,effective amounts will depend on the particular condition being treatedand the desired outcome. A therapeutically effective dose may bedetermined by those of ordinary skill in the art, for instance,employing factors such as those described herein and using no more thanroutine experimentation.

In certain embodiments of the invention, the administration of thecomposition of the invention may be designed so as to result insequential exposures to the composition over a certain time period, forexample, hours, days, weeks, months, or years. This may be accomplished,for example, by repeated administrations of a composition of theinvention by one or more of the methods described herein, or by asustained or controlled release delivery system in which the compositionis delivered over a prolonged period without repeated administrations.Administration of the composition using such a delivery system may be,for example, by a transdermal patch. Maintaining a substantiallyconstant concentration of the composition may be preferred in somecases.

For certain chronic treatments or therapies, it is contemplated that acomposition as discussed herein may be used to deliver nitric oxide tothe skin or mucosal surface at a relatively high concentration during aninitial treatment, and the amount of nitric oxide may be lowered or“titrated” down to a relatively lower concentration maintenance dose oramount. A nitric oxide containing composition as described herein can beused to promote vasodilation of blood vessels within and/or under theskin. As a specific example, a composition described herein may be usedfor certain treatments of hair loss or wounds, e.g., anal fissures. Insome cases, gradual improvement may be observed with successiveapplications.

In one set of embodiments, a composition such as is discussed herein maybe applied to the skin or mucosal surface of a subject, e.g., at anysuitable location. The composition may be contacted using any suitablemethod. For example, the composition may be rubbed on, poured on,applied with an applicator (e.g., a gauze pad, a swab, a bandage, etc.),or the like. In some cases, the composition can be a liquid, a gel, acream, a lotion, an ointment, a solid “stick,” or the like, that can beapplied to the skin or mucosal surface by hand, for example, by rubbingor spraying.

In certain embodiments, the composition is applied to a mucosal surfaceof the subject. For example, the composition may be applied to the noseor nostrils, the mouth, the lips, the eyelids, the ears, the genitalarea (of either male or female subjects), or the anus.

The composition may be applied, in certain embodiments, at or near agenital region of the skin of a male or a female subject, e.g., to treatsexual dysfunction. For example, the composition may be applied to thepenis of a male subject or to the vulva of a female subject, or to anyother suitable genital perianal region. In certain embodiments, forinstance, the composition is used to treat erectile dysfunction. In somecases, the composition can be applied to a condom or other suitablesexual aid. The composition can be applied, for example, just before thesubject engages in sexual activity.

Compared to other means of administration, the use of topical (ormucosal) administration in certain embodiments of the present inventionhas various advantages, including one or more of the following. In somecases, administration of a composition and delivery of nitric oxide asdiscussed herein is easier and more effective than other drugadministration routes, for example, oral delivery. Unlike oraladministration where a substantial amount of nitric oxide may bedestroyed during the digestive process, nitric oxide delivered topicallyor to a mucosal surface is not exposed to the digestive tract. Topicalor mucosal application may also allow, in some instances, relativelysteady delivery of nitric oxide to the desired target area without thecyclic dosages typical of orally or parenterally administered drugs. Insome embodiments, topical or mucosal application may also avoid toxicside effects associated with sustained increased levels of nitric oxidetypical of oral or parenteral administration.

Compared to other topical or mucosal delivery systems that employ nitricoxide donors (an entity that is able to release nitric oxide, such asL-arginine, nitroglycerin, or amyl nitrite) as a nitric oxide source,various aspects of the present invention utilizing nitric oxide gas haveseveral advantages, including one or more of the following. Nitric oxidecan be released relatively quickly in some embodiments, because therelease does not necessarily involve chemical transformations of nitricoxide donors to release nitric oxide. The concentration of nitric oxidecan accumulate quickly upon topical (or mucosal) administration, leadingto good therapeutic effect in certain embodiments of the invention.Thus, for example, rapid nitric oxide delivery may provide quickereffect in addressing issues of erectile dysfunction (or other sexualdysfunction in either male or female subjects) than is possible withoral dosage forms such as sildenafil. In other applications, it isexpected that nitric oxide can be similarly delivered rapidly. In someembodiments, the release rate of nitric oxide can be controlled, forinstance, by physical actions (e.g., by controlling how much of thecomposition is applied to the skin), in comparison to nitric oxidedonors which release nitric oxide upon chemical stimulation. Moreover,certain embodiments of the present invention employ phosphatidylcholine,a component of cell membranes, as a carrier which improves thepenetration and absorption of nitric oxide into cells and tissues. Thus,certain compositions of the present invention will be non-toxic orbiocompatible.

The compositions of the present invention may additionally comprise oneor more adjunct ingredients, for instance, pharmaceutical drugs or skincare agents. For example, compositions of the invention may includeadjuvants such as salts, buffering agents, diluents, excipients,chelating agents, fillers, drying agents, antioxidants, antimicrobials,preservatives, binding agents, bulking agents, silicas, solubilizers, orstabilizers. Non-limiting examples include species such as calciumcarbonate, sodium carbonate, lactose, kaolin, calcium phosphate, orsodium phosphate; granulating and disintegrating agents such as cornstarch or algenic acid; binding agents such as starch, gelatin oracacia; lubricating agents such as magnesium stearate, stearic acid, ortalc; time-delay materials such as glycerol monostearate or glyceroldistearate; suspending agents such as sodium carboxymethylcellulose,methylcellulose, hydroxypropylmethylcellulose, sodium alginate,polyvinylpyrrolidone; dispersing or wetting agents such as lecithin orother naturally-occurring phosphatides; thickening agents such as cetylalcohol or beeswax; buffering agents such as acetic acid and saltsthereof, citric acid and salts thereof, boric acid and salts thereof, orphosphoric acid and salts thereof; or preservatives such as benzalkoniumchloride, chlorobutanol, parabens, or thimerosal. Suitableconcentrations can be determined by those of ordinary skill in the art,using no more than routine experimentation. Those of ordinary skill inthe art will know of other suitable formulation ingredients, or will beable to ascertain such, using only routine experimentation.

Preparations can include sterile aqueous or nonaqueous solutions,suspensions and emulsions, which can be isotonic with the blood of thesubject in certain embodiments. Examples of nonaqueous solvents arepolypropylene glycol, polyethylene glycol, vegetable oil such as oliveoil, sesame oil, coconut oil, arachis oil, peanut oil, mineral oil,organic esters such as ethyl oleate, or fixed oils including syntheticmono or di-glycerides. Aqueous solvents include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution,1,3-butandiol, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's or fixed oils. Intravenous vehicles include fluid and nutrientreplenishers, electrolyte replenishers (such as those based on Ringer'sdextrose), and the like. Preservatives and other additives may also bepresent such as, for example, antimicrobials, antioxidants, chelatingagents and inert gases and the like. Those of skill in the art canreadily determine the various parameters for preparing and formulatingthe compositions of the invention without resort to undueexperimentation.

In another aspect, the present invention is directed to a kit includingone or more of the compositions discussed herein. A “kit,” as usedherein, typically defines a package or an assembly including one or moreof the compositions of the invention, and/or other compositionsassociated with the invention, for example, as described herein. Each ofthe compositions of the kit may be provided in liquid form (e.g., insolution), or in solid form (e.g., a dried powder). In certain cases,some of the compositions may be constitutable or otherwise proces sable(e.g., to an active form), for example, by the addition of a suitablesolvent or other species, which may or may not be provided with the kit.Examples of other compositions or components associated with theinvention include, but are not limited to, solvents, surfactants,diluents, salts, buffers, chelating agents, fillers, antioxidants,binding agents, bulking agents, preservatives, drying agents,antimicrobials, needles, syringes, packaging materials, tubes, bottles,flasks, beakers, dishes, frits, filters, rings, clamps, wraps, patches,containers, and the like, for example, for using, administering,modifying, assembling, storing, packaging, preparing, mixing, diluting,and/or preserving the compositions components for a particular use, forexample, to a sample and/or a subject.

A kit of the invention may, in some cases, include instructions in anyform that are provided in connection with the compositions of theinvention in such a manner that one of ordinary skill in the art wouldrecognize that the instructions are to be associated with thecompositions of the invention. For instance, the instructions mayinclude instructions for the use, modification, mixing, diluting,preserving, administering, assembly, storage, packaging, and/orpreparation of the composition and/or other compositions associated withthe kit. In some cases, the instructions may also include instructionsfor the delivery and/or administration of the compositions, for example,for a particular use, e.g., to a sample and/or a subject. Theinstructions may be provided in any form recognizable by one of ordinaryskill in the art as a suitable vehicle for containing such instructions,for example, written or published, verbal, audible (e.g., telephonic),digital, optical, visual (e.g., videotape, DVD, etc.) or electroniccommunications (including Internet or web-based communications),provided in any manner.

U.S. Provisional Patent Application Ser. No. 61/453,836, filed Mar. 17,2011, entitled “Topical Nitric Oxide Systems,” by Nicholas V. Perriconeis incorporated herein by reference in its entirety.

The following examples are intended to illustrate certain embodiments ofthe present invention, but do not exemplify the full scope of theinvention.

EXAMPLE 1

This example illustrates one technique for preparing a composition inaccordance with one embodiment of the invention. An accurate amount of acarrier (HNC 167-62) (see below) was introduced into a system. Thecarrier weight used in these experiments was approximately 250 g and thevessel size was 500 ml. The vessel was equipped with a mechanicalstirrer, gas inlet, and gas outlet and was previously purged with argonfor about an hour. The temperature of the carrier was kept at about25-30° C. NO gas regulated at 5 psi (1 psi is about 6,900 Pa) and wasthen introduced at a controlled rate of about 1 bubble/s with continuousstirring. The color, consistency, and viscosity of the carrier did notappear to change if NO was bubbled for 30 minutes to 2 hours. After 6hours, the weight of the carrier had increased by 0.15%, by 12 hours by0.25%, and by 24 hours by 0.56%. These increases in weight were believedto be significant considering the relative small molecular weight of NOversus the carrier. Although there was a slight change in color duringthe experiment (the color changed to slightly more orange), IR spectrumanalysis of the final product did not show any change versus the initialcarrier, indicating no noticeable chemical change in the carrier. Thecarrier also can solidify upon cooling if the carrier is initially asolid at lower temperature. Accordingly, this example demonstrates thata composition containing NO can be prepared in accordance with oneembodiment of the invention.

EXAMPLE 2

In this example, six experiments were carried out to investigate theinteraction of nitric oxide with three carriers (HNC 157-62, HNC 157-65,and HNC 157-69) as well as with 1,3-propanediol, using experimentalconditions similar to that described for Example 1. In addition threeexperiments were performed to prepare carriers containing 800 ppm and500 ppm nitric oxide. HNC 157-62 was formed of 65% Phospholipon-90 G(American Lecithin Company), 18% isopropyl palmitate (Kraft Chemicals),8% capric caprylic triglycerides (RITA Corp.), and 9% propanediol(Dupont). HNC 157-65 was formed of 65% Phospholipon-90 G, 13% isopropylpalmitate, 14% capric caprylic triglycerides, 3% propanediol, and 5%dimethyl isosorbide (Croda). HNC 157-69 was formed from 65%Phospholipon-90 G, 16% isopropyl palmitate, and 19% capric caprylictriglycerides.

The compositions were generally prepared as follows. Isopropylpalmitate, capric caprylic triglyceride, propanediol (for HNC 157-62 andHNC 157-65), and dimethyl isosorbide (for HNC 157-65) were mixedtogether and warmed to 40° C. Phospholipon-90 G was then gradually addedto this liquid mixture by mixing it. Phospholipon-90 G is typicallyreceived as individual pellets, and is mixed into the solution untilfully dissolved. The mixture was subsequently filtered through a sieveto remove any undissolved Phospholipon-90 G.

Accordingly, the HNC carriers included 1,3-propanediol, Phospholipon-90G, isopropyl palmitate, capric and/or caporic triglycerides, andArlasolve DMI (ICI America or Croda). Isopropyl palmitate, the capricand/or caporic triglycerides, and Arlasolve DMI are expected to bechemically inert towards nitric oxide, while the literature suggeststhat 1,2-propanediol and glycerol may be able to react with nitric oxidegas to form mononitrates. Accordingly, it would be expected that1,3-propanediol may also react with NO to form mononitrates:

In addition, Phospholipon-90 G is derived from soybean and containsesters of unsaturated fatty acids such as oleic, linoleic, and linolenicacids, and thus, the unsaturated fatty acid part of Phospholipon-90Gwould react with nitric oxide to lead to a variety of nitrated products.

Each carrier was taken in a 500 mL three necked flask equipped with amechanical stirrer, gas inlet and a gas outlet. The system was purgedwith argon for one hour at room temperature (25° C.). Then nitric oxidegas was bubbled into the system. Then, nitric oxide gas was bubbledthrough carrier for stipulated amount of time. The changes in weight andcolor were noted. The details of individual experiments were as follows.

Experiment 1. The carrier was HNC 157-62. Nitric oxide gas was bubbledfor 24 hours at 25° C. The initial weight of carrier was 168.53 g., andthe final weight was 169.48 g. The net weight gained was 0.95 g and thepercentage weight gain was 0.56%.

Experiment 2. The carrier used was HNC 157-62. Nitric oxide gas wasbubbled for 48 hours at 25° C. The initial weight of carrier was 171.31g., and the final weight was 174.21 g. The net weight gained was 2.90Gand the percentage weight gain was 1.69%.

Experiment 3. In order to differentiate between chemical reaction vs.physical absorption, the above reaction mixtures were heated at 55-60°C. for four hours. Minimal loss of weight was observed (˜200 mg),indicating no loss of absorbed nitric oxide gas. However, more intenseorange color developed during this process, indicating somedecomposition of the nitrites formed.

Experiment 4. The carrier used was HNC 157-65. Nitric oxide gas wasbubbled for 24 hours at 25 ° C. The initial weight of carrier was 171.66g., and the final weight was 172.98 g. The net weight gained was 1.32 gand percentage weight gain was 0.77%.

Experiment 5. The carrier used was HNC 157-69 (same as HNC 157-62,except it had no 1,3-propanediol). Nitric oxide gas was bubbled for 40hours at 25° C. The initial weight of carrier was 171.02 g., and thefinal weight was 171.97 g. The net weight gained was 0.95 g and thepercentage weight gain was 0.56%.

Experiment 6. Nitric oxide gas was bubbled through 1,3-propanediol(neat) for 40 hours at 25° C. The initial weight of the 1,3-propanediolwas 178.81 g., and the final weight was 178.97 g. The net weight gainedwas 0.16 g and the percentage weight gain was 0.09%.

Experiment 7. For preparation of 800 ppm NO, the carrier used was HNC157-62. Nitric oxide gas was bubbled for 2 hours at 25° C. The initialweight of carrier was 238.16 g., and the final weight was 238.35 g. Thenet weight gained was 0.19 g and the percentage weight gain 0.0798%(˜800 ppm). See entry 5 in Table 1.

Experiment 8. For preparation of 500 ppm NO, the carrier used was HNC157-65. Nitric oxide gas was bubbled for 2 hours at 25° C. The initialweight of carrier was 250.37 g., and the final weight was 250.50 g. Thenet weight gained was 0.13 g and the percentage weight gain was 0.0519%(˜500 ppm). See entry 6 in Table 1.

Experiment 9. For preparation of 800 ppm NO, the carrier used was HNC157-62. Nitric oxide gas was bubbled for 15 min at 25° C. The initialweight of carrier was 252.24 g., and the final weight was 252.45 g. Thenet weight gained was 0.21 g and the percentage weight gain 0.083% (˜800ppm).

These experiments were conducted with carriers the HNC 157-62, HNC157-65, HNC 157-69, and 1,3-propanediol.

As described above and in Table 1, weight gains ranging from 0.5% to1.7% were observed when nitric oxide gas was passed through thecarriers. In order to determine the nature of interaction between nitricoxide and carrier, the carrier was heated after nitric oxide absorptionat 60° C. for four hours. Practically no loss of weight was observed,which indicated that the nitric oxide gas reacted chemically with thecarriers (entries 1-4 in Table 1).

In order to investigate the reactivity of 1,3-propanediol with nitricoxide, nitric oxide absorption was studied using (a) HNC 157-69, whichdid not contain 1,3-propanediol, and (b) 1,3-propanediol by itself. HNC157-69 gained 0.95 g or 0.56% weight, much lower compared to its1,3-propanediol containing analog HNC 157-62, which showed 1.69% weightgain (entries 2 and 5 of Table 1). 1,3-propanediol itself, surprisingly,showed only negligible, if any, weight gain when NO was passed throughit (entry 6 in Table 1). Thus, under experimental conditions,1,3-propanediol did not react with nitric oxide.

Two samples were also prepared containing 800 ppm NO (from carrier HNC157-62) and one sample containing 500 ppm NO (from carrier HNC 157-65)(entries 7-9 in Table 1). The IR spectra of the carriers did not showany additional bands after the reaction, possibly because of low amountsof nitrites and/or overlap with the carrier complex bands.

Mass spectral studies of the carrier HNC 157-62 and HNC 157-62containing NO indicated that there was an increase in the intensity ofthe peak at m/e 104 in NO-containing carrier, compared to carrierwithout NO. The peak at m/e 104 was believed to be due to choline cation(C₅H₁₄NO). Phospholipon-90 G may contain some free choline, and hencepresence of the peak at 104 in the mass spectrum of the carrier was notsurprising. However, the increase in the amount of choline after passageof NO was somewhat unexpected, although it is believed that nitric oxidecatalyzes similar dephosphorylation of Phospholipon-90 G releasingcholine.

In conclusion, an increase in weight (0.56 to 1.69%) was observed whennitric oxide gas was passed through the carriers. 1,3-propanediol failedto gain any significant weight when nitric oxide was passed through it.HNC 157-69 (devoid of 1,3-propanediol) gained only 0.56% weight comparedto 1.69% by its 1,3-propanediol containing analog HNC 157-62. The massspectra of HNC 157-62 before and after passing NO indicated that thepeak corresponding to choline at m/e 104 increased after the passage ofNO, which suggests that phospholipon-90 G may undergo NO-catalyzeddephosphorylation.

TABLE 1 Initial wt. Final wt. Time Temp. % Wt. Expt. No. Carrier g. g.hr ° C. Wt. Gain g gain 1 HNC 157- 168.53 169.48 24 25 0.95 0.56 62 2HNC 171.31 174.21 48 25 2.90 1.69 157-62 3 HNC 174.21* 174.01 4 60 −0.20−0.11 157-62 4 HNC 171.66 172.98 24 25 1.32 0.77 157-65 5 HNC 171.02171.97 40 25 0.95 0.56 157-69 6 1,3- 178.81 178.97 40 25 0.16 0.09Propanediol 7 HNC 238.16 238.35 2 25 0.19 0.0798 157-62 (~800 ppm) 8 HNC250.37 250.50 2 25 0.13 0.0519 157-65 (~500 ppm) 9 HNC 252.24 252.450.25 25 0.21 0.0833 157-62 (~800 ppm)

While several embodiments of the present invention have been describedand illustrated herein, those of ordinary skill in the art will readilyenvision a variety of other means and/or structures for performing thefunctions and/or obtaining the results and/or one or more of theadvantages described herein, and each of such variations and/ormodifications is deemed to be within the scope of the present invention.More generally, those skilled in the art will readily appreciate thatall parameters, dimensions, materials, and configurations describedherein are meant to be exemplary and that the actual parameters,dimensions, materials, and/or configurations will depend upon thespecific application or applications for which the teachings of thepresent invention is/are used. Those skilled in the art will recognize,or be able to ascertain using no more than routine experimentation, manyequivalents to the specific embodiments of the invention describedherein. It is, therefore, to be understood that the foregoingembodiments are presented by way of example only and that, within thescope of the appended claims and equivalents thereto, the invention maybe practiced otherwise than as specifically described and claimed. Thepresent invention is directed to each individual feature, system,article, material, kit, and/or method described herein. In addition, anycombination of two or more such features, systems, articles, materials,kits, and/or methods, if such features, systems, articles, materials,kits, and/or methods are not mutually inconsistent, is included withinthe scope of the present invention.

All definitions, as defined and used herein, should be understood tocontrol over dictionary definitions, definitions in documentsincorporated by reference, and/or ordinary meanings of the definedterms.

The indefinite articles “a” and “an,” as used herein in thespecification and in the claims, unless clearly indicated to thecontrary, should be understood to mean “at least one.” The phrase“and/or,” as used herein in the specification and in the claims, shouldbe understood to mean “either or both” of the elements so conjoined,i.e., elements that are conjunctively present in some cases anddisjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e., “one or more” ofthe elements so conjoined. Other elements may optionally be presentother than the elements specifically identified by the “and/or” clause,whether related or unrelated to those elements specifically identified.Thus, as a non-limiting example, a reference to “A and/or B”, when usedin conjunction with open-ended language such as “comprising” can refer,in one embodiment, to A only (optionally including elements other thanB); in another embodiment, to B only (optionally including elementsother than A); in yet another embodiment, to both A and B (optionallyincluding other elements); etc.

As used herein in the specification and in the claims, “or” should beunderstood to have the same meaning as “and/or” as defined above. Forexample, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items. Only terms clearly indicated tothe contrary, such as “only one of” or “exactly one of,” or, when usedin the claims, “consisting of,” will refer to the inclusion of exactlyone element of a number or list of elements. In general, the term “or”as used herein shall only be interpreted as indicating exclusivealternatives (i.e. “one or the other but not both”) when preceded byterms of exclusivity, such as “either,” “one of,” “only one of,” or“exactly one of.” “Consisting essentially of,” when used in the claims,shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase “atleast one,” in reference to a list of one or more elements, should beunderstood to mean at least one element selected from any one or more ofthe elements in the list of elements, but not necessarily including atleast one of each and every element specifically listed within the listof elements and not excluding any combinations of elements in the listof elements. This definition also allows that elements may optionally bepresent other than the elements specifically identified within the listof elements to which the phrase “at least one” refers, whether relatedor unrelated to those elements specifically identified. Thus, as anon-limiting example, “at least one of A and B” (or, equivalently, “atleast one of A or B,” or, equivalently “at least one of A and/or B”) canrefer, in one embodiment, to at least one, optionally including morethan one, A, with no B present (and optionally including elements otherthan B); in another embodiment, to at least one, optionally includingmore than one, B, with no A present (and optionally including elementsother than A); in yet another embodiment, to at least one, optionallyincluding more than one, A, and at least one, optionally including morethan one, B (and optionally including other elements); etc.

It should also be understood that, unless clearly indicated to thecontrary, in any methods claimed herein that include more than one stepor act, the order of the steps or acts of the method is not necessarilylimited to the order in which the steps or acts of the method arerecited.

In the claims, as well as in the specification above, all transitionalphrases such as “comprising,” “including,” “carrying,” “having,”“containing,” “involving,” “holding,” “composed of,” and the like are tobe understood to be open-ended, i.e., to mean including but not limitedto. Only the transitional phrases “consisting of” and “consistingessentially of” shall be closed or semi-closed transitional phrases,respectively, as set forth in the United States Patent Office Manual ofPatent Examining Procedures, Section 2111.03.

What is claimed is: 1-192. (canceled)
 193. A composition, comprising: anemulsion comprising a first phase comprising nitric oxide and a carriercomprising lecithin containing the nitric oxide, and a second phasecomprising an emulsifier, wherein the lecithin is present at at leastabout 0.25% by weight of the composition, and wherein the composition isstable at room temperature.
 194. The composition of claim 193, whereinthe first phase forms discrete vesicles contained within the secondphase.
 195. The composition of claim 193, wherein the nitric oxide ispresent at at least about 0.5% by weight of the composition.
 196. Thecomposition of claim 193, wherein the lecithin comprises aphosphatidylcholine.
 197. The composition of claim 196, wherein thephosphatidylcholine is present at between about 8% and about 65% byweight of the composition.
 198. The composition of claim 196, wherein atleast some of the phosphatidylcholine comprises apolyenylphosphatidylcholine.
 199. The composition of claim 193, whereinthe composition comprises no more than about 250 ppm of water by weightof the composition.
 200. The composition of claim 193, wherein the firstphase comprises a fatty acid ester.
 201. The composition of claim 193,wherein the composition is a gel.
 202. The composition of claim 193,wherein the composition is a cream.
 203. The composition of claim 193,wherein the composition is substantially transparent.
 204. Thecomposition of claim 193, wherein the composition has a viscosity of atleast about 20,000 cP.
 205. The composition of claim 193, wherein thenitric oxide is present within the emulsion as a gas or bound byhydrogen bonds or van der Waals forces to the lecithin.
 206. A method,comprising contacting the skin of a subject with the composition ofclaim
 193. 207. A method, comprising contacting a mucosal surface of asubject with the composition of claim
 193. 208. A method, comprisingcontacting the penis of a male subject with the composition of claim193.
 209. A method, comprising contacting the vulva of a female subjectwith the composition of claim
 193. 210. A method, comprising contactinga wound on the skin of a subject with the composition of claim
 193. 211.An article comprising a container containing the composition of claim193, wherein the container is sealed.
 212. An article comprising acontainer containing the composition of claim 193, wherein the containeris substantially watertight.